Nutritional folate-deficiency in Chinese hamster ovary cells: Chromosomal abnormalities associated with perturbations in nucleic acid precursors

Abstract

Folate deficiency is known to induce chromosomal abnormalities. We used a nutritionally folate-deficient Chinese hamster ovary (CHO) cell culture system to examine modulation of chromosome damage by purine or pyrimidine supplementation. The cells were cultured in folate-deficient (Fol −) medium or Fol − medium supplemented with thymidine (dT) or hypoxanthine (Hx) until population growth arrest. The cultures were then switched to complete medium, permitting the cells to begin cell division. Cell-cycle progression was followed by flow cytometry to identify the first mitosis, when samples for analysis were collected. The mitotic index, frequency of chromosomal aberrations in mitotic cells, and relative distribution of different types of aberrations were determined. Cells grown in Fol − medium supplemented with Hx entered the G 2/M phase of the cell cycle at 14 hours after media change as compared with 16 hours for Fol − cultures or 24 hours for Fol − cultures supplemented with dT. Cells cultured in Fol − medium alone or supplemented with dT showed similar frequencies of damage, averaging 20–22%, as compared with 2% for control cultures. In contrast, cells grown in Hx-supplemented medium exhibited a lower frequency of damaged mitoses (15%), as well as a reduction in certain types of abnormalities. This latter finding is surprising in light of our previous work showing that the presence of Hx during folate deficiency produced a more severe perturbation of phenotype (cellular enlargement) and growth control (S-phase delay and cell killing) than did dT supplementation or the absence of both nucleotide precursors.